ES2914072T3 - Monocyclic pyridine derivative salt and its crystal - Google Patents

Abstract

A salt consisting of 5-({2-[({4-[1-(2-hydroxyethyl)piperidin-4-yl]phenyl}carbonyl)amino]pyridin-4-yl}oxy)-6-(2- methoxyethoxy)-N-methyl-1H-indole-1-carboxamide represented by formula (I): **(See formula)** and succinic acid or maleic acid.

Description

DESCRIPTION

Monocyclic pyridine derivative salt and its crystal

field of invention

The present invention relates to a monocyclic pyridine derivative salt and a crystal thereof having an FGFR inhibitory action.

State of the art

An FGF (fibroblast growth factor) is known as a growth factor to control a variety of physiological functions such as cell growth, cell migration, cell infiltration, cell survival, differential induction, wound healing and the angiogenesis.

FGF controls various physiological functions via FGF receptors (FGFRs: FGFR1, FGFR2, FGFR3, and FGFR4), ie, receptor tyrosine kinases. Each FGFR includes three types of domains: an extracellular domain, a transmembrane domain, and an intracellular tyrosine kinase domain. When an FGF binds to the extracellular domain of an FGFR, a receptor dimer is formed. Subsequently, the intracellular tyrosine kinase is activated, and then, an intracellular signal is transmitted mainly through an MPAK (mitogen-activated protein kinase)/ERF (extracellular signal-regulated kinase) mechanism or a PI3K (phosphatidylinositol 3) mechanism. -kinase)/Akt.

At the same time, it has been reported that various types of cancer such as breast cancer, bladder cancer, EMS (8p11 myeloproliferative syndrome), stomach cancer, endometrial cancer and prostate cancer are caused as a result of the induction of abnormality of FGF/FGFR signal accompanying enhancement of FGF production, FGFR gene amplification, FGFR overexpression, FGFR fusion protein production, FGFR mutation (Non-patent Literature 1). In addition, the following have been reported as cancers accompanied by the FGF/FGFR signal abnormality: non-small cell lung carcinoma, small cell lung carcinoma, ovarian cancer, sarcoma, colon cancer, melanoma, glioblastoma, astrocytoma, head and neck cancer (Non-Patent Literature 2 and 3), thyroid cancer (Non-Patent Literature 4), pancreatic cancer (Non-Patent Literature 5 and 6), liver cancer (Non-Patent Literature 7), skin cancer (Non-Patent Literature 8), kidney cancer (Non-Patent Literature 9), and squamous cell carcinoma of the lung (Non-Patent Literature 10, 11 and 12).

In addition, the FGF/FGFR signal is one of the main angiogenic signals in endothelial cells along with a VEGF (vascular endothelial growth factor)/KDR (insert kinase domain-containing receptor) signal, and is reported to be involved in the interaction between cancer stromal cells (fibroblasts) and cancer cells (Non-Patent Literature 1).

Therefore, an FGFR inhibitor that directs an FGF/FGFR signal is expected to function as an antitumor drug, against cancers accompanied by the FGF/FGFR signal abnormality, based on its inhibitory action against the signal abnormality and its action. inhibitory against the angiogenic signal. Recently, a selective FGFR inhibitor referred to as insusceptible to be affected by the confrontation effect of another signal, such as a selective FGFR inhibitor against FGFR1, FGFR2 or FGFR3, which is obviously different in structure from a compound of the present invention. In human antitumor drug development, however, the selective FGFR inhibitor is beyond an antitumor drug that simultaneously targets both FGF/FGFR signal and VEGF/KDR signal and has not been put into practice. the market yet (Non-Patent Literatures 13 and 14; Non-Patent Literatures 1 and 2. Non-Patent Literature 3 discloses pyrimidine derivatives but does not disclose an inhibitory action against FGF/FGFR signal abnormality. Non-patent literature 4 discloses pyridine derivatives or pyrimidine derivatives that inhibit VEGF- and FGF-induced angiogenesis None of these literatures, however, disclose the compounds of the present invention.

citation list

Patent bibliography

Patent Literature Reference 1: International Publication No. WO 2008/075068.

Patent Literature Reference 2: International Publication No. WO 2006/000420.

Patent Literature Reference 3: International Publication No. WO 2002/032872.

Patent Literature Reference 4: International Publication No. WO 2004/020434.

Non-patent literature

Non-patent literature reference 1: Nicholas et al., "Fibroblast growth factor signaling: from development to cancer," Nature Reviews Cancer. 2010; 10: 116-129

Non-patent literature reference 2: Jorgen WESCHE et al., Fibroblast growth factors and their receptors in cancer, Biochem J. 2011: 437; 199-213

Non-patent literature reference 3: Gennaro Daniele et al., FGF Receptor Inhibitors: Role in Cancer Therapy, Curr Oncol Rep. 2012; 14:111-119

Non-patent literature reference 4: Rosanne St. Bernard et al., Fibroblast Growth Factor Receptors as Molecular Targets in Thyroid Carcinoma, Endocrinology. 2005; 146: 1145-1153

Non-patent literature reference 5: Toshiyuki Ishiwata et al., Enhanced Expression of Fibroblast Growth Factor Receptor 2 IIIc Promotes Human Pancreatic Cancer Cell Proliferation, Am J Pathol. 2012; 180: 1928-1941

Non-patent literature reference 6: G Chen et al., Inhibition of endogenous SPARC enhances pancreatic cancer cell growth: modulation by FGFR1-III isoform expression, Br J Cancer. 2010; 102: 188-195 Non-patent literature reference 7: Dorothy M. French et al., Targeting FGFR4 Inhibits Hepatocellular Carcinoma in Preclinical Mouse Models, PLoS One. 2012; 7: e36713

Non-patent literature reference 8: Armelle Logie et al., Activating mutations of the tyrosine kinase receptor FGFR3 are associated with benign skin tumors in mice and humans, Hum Mol Genet 2005; 14: 1153 1160

Non-patent literature reference 9: Tsimafeyeu I et al., Overexpression of fibroblast growth factor receptors FGFR1 and FGFR2 in renal cell carcinoma, Scand J Urol Nephrol 2011; 45: 190-195 Non-patent literature reference 10: Jonathan Weiss et al., Frequent and Focal FGFR1 Amplification Associates with Therapeutically Tractable FGFR1 Dependency in Squamous Cell Lung Cancer, Sci Transl Med. 2010; 2: issue 6262-93

Non-patent literature reference 11: Hidefumi Sasaki et al., Increased FGFR1 copy number in lung squamous cell carcinomas, Mol Med Report. 2012; 5: 725-728

Non-patent literature reference 12: The Cancer Genome Atlas Research Network, Comprehensive genomic characterization of squamous cell lung cancers, Nature 2012; 489: 519-525 Non-patent literature reference 13: Paul R Gavine et al., AZD4547: An Orally Bioavailable, Potent, and Selective Inhibitor of the Fibroblast Growth Factor Receptor Tyrosine Kinase Family, Cancer Res. 2012; 72: 2045-2056

Non-patent literature reference 14: Vito Guagnano et al., Discovery of 3-(2,6-Dichloro -3,5-dimethoxy-phenyl)-1 -{6-[4-(4-ethyl-piperazin -1 -yl)-phenylamino]-pyrimidin-4-yl}-1 -methyl-urea (NVP-BGJ398), A Potent and Selective Inhibitor of the Fibroblast Growth Factor Receptor Family of Receptor Tyrosine Kinase, J Med Chem. 2011; 54: 7066-7083

Summary of the invention

technical problem

A compound represented by the following formula (I) 5-({2-[({4-[1-(2-hydroxyethyl)piperidin-4-yl]phenyl}carbonyl)amino]pyridin-4-yl}oxy )-6-(2-methoxyethoxy)-N-methyl-1H-indole-1-carboxamide, hereinafter referred to as compound (I)) has FGFR1, FGFR2 and FGFR3 inhibitory actions. Generally, the physical properties of a compound, one of its salts, and its crystals used as a pharmaceutical greatly affect the bioavailability of a drug, the purity of the active pharmaceutical ingredient, and/or the prescription of a preparation. An object of the present invention is therefore to provide a salt of a compound (I) or a crystal thereof, which can be used as raw materials for pharmaceutical substances.

The present inventors have carried out earnest studies on a compound (I) in consideration of the above-mentioned circumstances, and as a result, have discovered salts of compound (I) or their crystals, thus completing the invention.

Solution to the problem

Specifically, the present invention provides the following [1] to [12].

[1] A salt consisting of 5-({2-[({4-[1-(2-hydroxyethyl)piperidin-4-yl]phenyl}carbonyl)amino]pyridin-4-yl}oxy)-6- (2-methoxyethoxy)-N-methyl-1H-indole-1-carboxamide represented by formula (I):

and succinic acid or maleic acid.

[2] The salt according to [1] above, which is a succinate salt.

[3] The salt according to [2] above, which is 5-({2-[({4-[1-(2-hydroxyethyl)piperidin-4-yl]phenyl}carbonyl)amino]pyridine succinate salt 1.5 -4-yl}oxy)-6-(2-methoxyethoxy)-N-methyl-1 H -indole-1-carboxamide.

[4] The salt according to [2] above, which is 0.5 5-({2-[({4-[1-(2-hydroxyethyl)piperidin-4-yl]phenyl}carbonyl)amino]pyridine succinate salt -4-yl}oxy)-6-(2-methoxyethoxy)-N-methyl-1 H -indole-1-carboxamide.

[5] The salt according to [1] above, which is 5-({2-[({4-[1-(2-hydroxyethyl)piperidin-4-yl]phenyl}carbonyl)amino]pyridin- 4-yl}oxy)-6-(2-methoxyethoxy)-N-methyl-1 H -indole-1-carboxamide.

[6] A crystal of 5-({2-[({4-[1-(2-hydroxyethyl)piperidin-4-yl]phenyl}carbonyl)amino]pyridin-4-yl}oxy) succinate salt -6-(2-methoxyethoxy)-N-methyl-1H-indole-1-carboxamide, which has diffraction peaks at diffraction angles (20 ± 0.2°) of 22.4°, 25.3° and 23.3° in beam diffraction- X in powder form measured with copper irradiation; detector: scintillation counter; tube voltage: 50kV; tube current: 300mA; Slit: 0.5mm divergence slit (2mm height limitation slit), open spreading slit, open receiving slit; sweep speed: 10°/min; sampling interval: 0.02°; sweep range: 5° to 35°; sample holder made of aluminium.

[7] A crystal (a) of 0.5 5-({2-[({4-[1-(2-hydroxyethyl)piperidin-4-yl]phenyl}carbonyl)amino]pyridin-4 succinate salt -yl}oxy)-6-(2-methoxyethoxy)-N-methyl-1H-indole-1-carboxamide, having diffraction peaks at diffraction angles (20 ± 0.2°) of 19.8°, 15.7° and 13.9 ° in X-ray powder diffraction measured with copper irradiation; detector: scintillation counter; tube voltage: 50kV; tube current: 300mA; slit: 0.5mm divergence slit (2mm height limitation slit), open spreading slit, open receiving slit; sweep speed: 10°/min; sampling interval: 0.02°; sweep range: 5° to 35°; sample holder made of aluminium.

[8] A crystal of 5-({2-[({4-[1-(2-hydroxyethyl)piperidin-4-yl]phenyl}carbonyl)amino]pyridin-4-yl}oxy)- 6-(2-methoxyethoxy)-N-methyl-1H-indole-1-carboxamide, which has diffraction peaks at (26 ± 0.2°) diffraction angles of 20.1°, 17.0° and 16.2° in X-ray diffraction X in powder form measured with copper irradiation; detector: scintillation counter; tube voltage: 50kV; tube current: 300mA; Slit: 0.5mm divergence slit (2mm height limitation slit), open spreading slit, open receiving slit; sweep speed: 10°/min; sampling interval: 0.02°; sweep range: 5° to 35°; sample holder made of aluminium.

[9] A crystal of 5-({2-[({4-[1-(2-hydroxyethyl)piperidin-4-yl]phenyl}carbonyl)amino]pyridin-4-yl}oxy) succinate salt -6-(2-methoxyethoxy)-N-methyl-1H-indole-1-carboxamide, having peaks at chemical shifts (±0.5 ppm) of 108.5 ppm, 155.1 ppm and 179.9 ppm in a solid state NMR spectrum of 13C measured using the equipment and measurement conditions as described in the description.

[10] The crystal according to claim 9, having peaks at chemical shifts (±0.5 ppm) of 27.1 ppm, 34.8 ppm, 108.5 ppm, 155.1 ppm and 179.9 ppm in a 13C solid state NMR spectrum measured using the equipment and measurement conditions as described in the description.

[11] A crystal of 5-({2-[({4-[1-(2-hydroxyethyl)piperidin-4-yl]phenyl}carbonyl)amino]pyridin-4-yl}oxy) succinate salt -6-(2-methoxyethoxy)-N-methyl-1H-indole-1-carboxamide, having an X-ray diffraction pattern measured with copper irradiation; detector: scintillation counter; tube voltage: 50kV; tube current: 300mA; Slit: 0.5mm divergence slit (2mm height limitation slit), open spreading slit, open receiving slit; sweep speed: 10°/min; sampling interval: 0.02°; sweep range: 5° to 35°; sample holder made of aluminum as shown in Figure 1.

[12] A pharmaceutical composition comprising the salt or the crystal according to any one of [1 ]-[11 ] as an active ingredient.

Advantageous effects of the invention

The salts of compound (I) and its crystals provided by the present invention possess properties as shown in the examples, hygroscopic nature as shown in the test examples described below, and potential for use as a drug substance in pharmaceutical substances.

Brief description of the figures

Figure 1 is an X-ray powder diffraction pattern of the crystal of 1.5 succinate salt of compound (I) obtained in Example 1. The abscissa shows the diffraction angle (26) and the ordinate shows the intensity of the peak.

Figure 2 is an X-ray powder diffraction pattern of the crystal of 0.5 succinate salt of compound (I) (a) obtained in Example 2. The abscissa shows the diffraction angle (26) and the ordinate shows the intensity of the peak.

Figure 3 is an X-ray powder diffraction pattern of the crystal of the maleate salt of compound (I) obtained in Example 3. The abscissa shows the diffraction angle (26) and the ordinate shows the intensity of the peak. Figure 4 is a 13C solid state NMR spectrum of the crystal of 1.5 succinate salt of compound (I) obtained in Example 1.

Figure 5 is a pattern of hygroscopic nature of the crystal of 1.5 succinate salt of compound (I) obtained in Example 1. The abscissa shows the relative humidity and the ordinate shows the weight change.

Figure 6 is an X-ray diffraction pattern of the crystal of the free form of compound (I) (Free Form A) obtained in Preparation Example 1-15. The abscissa shows the diffraction angle (26) and the ordinate shows the intensity of the peak.

Figure 7 is an X-ray diffraction pattern of the crystal of the free form of compound (I) (Free Form B) obtained in Reference Example 1. The abscissa shows the diffraction angle (26) and the ordinate shows the intensity of the peak.

Figure 8 is an X-ray diffraction pattern of the free form crystal of compound (I) (Free Form Hydrate) obtained in Reference Example 2. The abscissa shows the diffraction angle (26) and the ordinate shows the intensity of the peak.

Figure 9 is an X-ray powder diffraction pattern of the mesylate salt crystal of compound (I) obtained in Reference Example 3. The abscissa shows the diffraction angle (26) and the ordinate shows the peak intensity.

Figure 10 is an X-ray powder diffraction pattern of the crystal of tosylate salt of compound (I) obtained in Reference Example 4. The abscissa shows the diffraction angle (26) and the ordinate shows the peak intensity.

Fig. 11 is an X-ray diffraction pattern of the benzoate salt crystal of compound (I) obtained in Reference Example 5. The abscissa shows the diffraction angle (26) and the ordinate shows the peak intensity. Fig. 12 is an X-ray diffraction pattern of the fumarate salt crystal of compound (I) obtained in Reference Example 6. The abscissa shows the diffraction angle (26) and the ordinate shows the peak intensity. Figure 13 is an X-ray powder diffraction pattern of the crystal of 0.5 succinate salt of compound (I) (a) obtained in Example 4. The abscissa shows the diffraction angle (26) and the ordinate shows the intensity of the peak.

Figure 14 is an X-ray powder diffraction pattern of the crystal of 0.5 succinate salt of compound (I) (b) obtained in Example 5. The abscissa shows the diffraction angle (26) and the ordinate shows the intensity of the peak.

Figure 15 is an X-ray powder diffraction pattern of the crystal of 1.5 succinate salt of compound (I) obtained in Example 6. The abscissa shows the diffraction angle (26) and the ordinate shows the intensity of the peak.

Description of achievements

A salt of the compound (I) of the present invention, a crystal thereof, and its production methods are described in detail below.

In the present description, a "salt" refers to a chemical entity consisting of the compound (I) as a basic component and a specific number of equivalents of an acid for the compound (I).

Examples of "salt" used herein include salts with succinic acid and maleic acid. And succinic acid is especially preferred.

The salts according to the present invention can be anhydrous, hydated or solvate. As used herein, a hydrate or solvate means that compound (I) or a salt thereof and water molecules or solvent molecules form together, and the solid may be crystalline. Examples of solvents in the solvates include ketone solvents such as acetone, 2-butanone, and cyclohexanone; ester solvents such as methyl acetate and ethyl acetate; ether solvents such as 1,2-dimethoxyethane and t-butyl methyl ether; alcohol solvents such as methanol, ethanol, 1-propanol, and isopropanol; and polar solvents such as N-methyl-2-pyrrolidone, N,N-dimethylformamide, and dimethylsulfoxide. The number of water molecules or solvent molecules with respect to the compound (I) or a salt thereof is not particularly limited, and examples thereof include one molecule or two molecules.

As used herein, a "crystal" refers to a crystal of a compound (I) or a salt thereof. Therefore, a crystal of 1.5 succinic acid salt of compound (I), for example, means a crystal of a salt formed between compound (I) and succinic acid, which has 1.5 molecules of succinic acid per 1 molecule of compound ( YO);

a crystal of 0.5 succinate salt (a) of a compound (I), which has diffraction peaks at diffraction angles (26 ± 0.2°) of 19.8°, 15.7° and 13.9° in X-ray diffraction as dust;

a crystal of maleate salt of a compound (I), which has diffraction peaks at diffraction angles (26 ± 0.2°) of 20.1°, 17.0° and 16.2° in X-ray powder diffraction;

a crystal of 1.5 succinate salt of a compound (I), characterized by having peaks at chemical shifts (±0.5 ppm) of 108.5 ppm, 155.1 ppm and 179.9 ppm in a 13C solid state NMR spectrum; either

a crystal of 1.5 succinate salt of compound (I) according to [9], characterized by having peaks at chemical shifts (±0.5 ppm) of 27.1 ppm, 34.8 ppm, 108.5 ppm, 155.1 ppm and 179.9 ppm in an NMR spectrum in 13C solid state.

The X-ray powder diffraction peaks described above are characteristic of the respective crystals and the diffraction peaks characteristic of the crystals of the 1.5 succinate salts of compound (I), 0.5 succinate salts (a) of compound (I) and maleate salts of compound (I).

In general, errors in diffraction angles (26) within the range of ± 0.2° can arise in X-ray powder diffraction and therefore the above described values of diffraction angles should be considered to include values within the range about ± 0.2°. Included in the present invention are not only crystals of certain salts with peaks at exactly the same diffraction angles in powder X-ray diffraction, but also crystals with peaks with an error range of approximately ± 0.2° from the diffraction angles.

Thus, for example, the phrase "having a diffraction peak at a diffraction angle (26 ± 0.2°) of 22.4°", as used herein, means "having a diffraction peak at an angle of diffraction (26) between 22.2° and 22.6°". The same applies to other diffraction angles as well.

Generally, the peak intensities and half-value widths of the diffraction angles (20) in X-ray powder diffraction are different for each measurement due to differences in the measurement conditions and the size and size dispersions. shape of each crystal particle in powder form and not always stable even though the crystal shapes are the same. Therefore, in the case of comparing an X-ray diffraction pattern, when the diffraction angles (20) are the same but the peak intensities and half-value widths are different, these differences do not imply that they come from differences in the shape of the crystal. Thus, a crystal having an X-ray diffraction pattern in powder form, having the aforementioned differences with respect to the characteristic diffraction peaks of a certain salt crystal according to the present invention, means that the crystal it has the same crystalline form as the crystal of the salt according to the present invention. As used herein, "having an X-ray powder diffraction pattern according to Fig. 1" means that it includes not only the case of having exactly the same pattern as shown in Fig. 1 , but also the case of having the same characteristic diffraction angles but different peak intensities and half-value widths. Thus, each crystal having such an X-ray diffraction pattern in powder form means that the crystal is identical to the crystal according to the present invention.

As used herein, "having peaks at chemical shifts (±0.5 ppm) of 27.1 ppm, 34.8 ppm, 108.5 ppm, 155.1 ppm, and 179.9 ppm" means having peaks each substantially equivalent to the peaks at chemical shifts (± 0.05 ppm) of 27.1 ppm, 34.8 ppm, 108.5 ppm, 155.1 ppm, and 179.9 ppm, when 13C solid-state NMR spectrometry is performed under the normal measurement condition or under a condition substantially the same as herein descriptive memory".

When determining whether or not it “has peaks substantially equivalent to” or not, the chemical shift values described above should be considered to include values within the range of approximately ± 0.5 ppm since errors in chemical shifts (ppm) can generally appear within of the ± 0.5 ppm range in a 13C solid state NMR spectrum. Included in the present invention, therefore, are not only crystals with exactly the same chemical shifts in a 13C solid state NMR spectrum, but also crystals with chemical shifts within an error range of about ± 0.5 ppm. Also, "having a peak at a chemical shift (±0.5 ppm) of 27.1 ppm" as used herein, for example, means "having a peak at a chemical shift of 26.6 ppm to 27.6 ppm." The same also applies to other chemical shifts in 13C solid state rMn spectra.

The production method of the salts of the compound (I) or its crystals, which are an embodiment according to the present invention, is illustrated below.

Production of Compound (I)

Compound (I) can be synthesized as specifically described in Production Example 1 below.

Methods for producing salts of compound (I)

The salts of the compound (I) according to the present invention can be obtained by means of conventional methods for producing salts. Specifically, they can be produced, for example, by suspending or dissolving the compound (I) in a solvent, heating if necessary, then adding an acid to the obtained suspension or solution, and stirring or leaving the solution. or resulting suspension for several minutes to several days at room temperature or with ice-bath cooling. The salts of compound (I) can be obtained as crystals or amorphous substances depending on the production methods. An amorphous substance can be prepared by adding the production methods to freeze-drying operations, if necessary. Examples of solvents to be used in these methods include alcohol solvents such as ethanol, 1-propanol, and isopropanol; acetonitrile; ketone solvents such as acetone and 2-butanone; ester solvents such as ethyl acetate; saturated hydrocarbon solvents such as hexane and heptane; ether solvents such as methyl t-butyl ether or water. Each of these solvents can be used alone, or two or more can be mixed and used.

Methods for producing crystals of compound (I) or its salts

A crystal of compound (I) or a salt thereof can be produced by the above-mentioned methods for producing compound (I) or a salt thereof, by thermal dissolution of compound (I) or a salt thereof in a solvent and crystallization thereof through cooling with stirring.

The compound (I) or a salt thereof that can be used in the crystallization may be in any form: it may be a solvate, a hydrate, an anhydride, an amorphous substance, a crystalline substance (including those consisting of a plurality of crystalline polymorphs) or one of their combinations.

Examples of solvents to be used in the crystallization include alcohol solvents such as methanol, ethanol, isopropanol, and 1-propanol; acetonitrile; amide solvents such as N,N-dimethylformamide; ester solvents such as ethyl acetate; saturated hydrocarbon solvents such as hexane and heptane; ketone solvents such as acetone and 2-butanone; ether solvents such as methyl t-butyl ether or water. Also, each of these solvents can be used alone, or two or more can be mixed and used.

The amount of the solvent to be used can be appropriately selected, provided that the lower limit is the amount in which the compound (I) or a salt thereof is obtained by heating or the suspension can be stirred, and The upper limit is the amount by which the performance of the crystal is not significantly reduced.

A seed crystal (eg, the crystal of the desired compound (I) or a salt thereof) may or may not be added during crystallization. The temperature, at which the crystal seed is added, is not particularly limited and is preferably 0 to 80°C.

The temperature to be employed when the compound (I) or a salt thereof is dissolved by heating, that at which the compound (I) or a salt thereof dissolves, can be appropriately selected depending on the solvent, but it is preferably within the range from 50°C to the temperature at which reflux of the crystallization solvent begins, and more preferably from 55 to 80°C.

Cooling during crystallization could yield substances containing different forms of crystals (polymorphism) if the cooling is rapid. Therefore, it is desirable to carry out cooling by controlling the cooling rate, as appropriate, taking into account its effect on crystal quality and/or grain size. For example, cooling with a cooling rate of between 5 and 40°C/hour is preferred. It is even more preferred that the cooling be at a cooling rate of, for example, between 5 and 25°C/hour.

Furthermore, the final crystallization temperature can be conveniently selected for crystal yield and/or quality, and is preferably between -25 and 30°C.

The desired crystal can be obtained by isolating the crystal formed by a conventional filtration method, washing the crystal separated by filtration with a solvent, if necessary, and then drying it. As the solvent to be used for washing the crystal, the same solvent as in crystallization can be used. Preferably, it is, for example, ethanol, acetone, 2-butanone, ethyl acetate, diethyl ether, methyl t-butyl ether and/or hexane. Each of these solvents can be used alone, or two or more can be mixed and used.

The crystal isolated by the filtration process may be dried, as appropriate, by exposure to air or a stream of nitrogen, or by heating.

As the drying time, the time until the residual solvent amount is less than the predefined amount can be selected, as appropriate, depending on the production amount, the drying apparatus and/or the drying temperature. Furthermore, the drying can be carried out with a current of air or under reduced pressure. The degree of pressure reduction can be selected, as appropriate, depending on the production amount, the drying apparatus and/or the drying temperature. The crystal obtained can be left in the air after drying if necessary.

The salts of compound (I) and its crystals can be formulated by a conventional method, and examples of dosage forms include oral formulations (such as tablets, granules, powders, capsules, and syrups), injections (for intravenous administration, intramuscular, subcutaneous administration and intraperitoneal administration) and external formulations (such as transdermal absorption formulations (such as ointments and patches), ophthalmic preparations, nasal preparations and suppositories).

To produce an oral solid formulation, a carrier, a binder, a disintegrant, a lubricant and/or a colorant, if necessary, can be added to the salts of the compound (I) and its crystals, and a tablet can be produced, a granule, a powder agent, or a capsule according to a conventional method. Furthermore, said tablet, granule, powder agent and/or capsule may be subjected to coating, if necessary.

Examples of the carrier include lactose, crystalline cellulose, examples of the binder include hydroxypropyl cellulose, examples of the disintegrant include croscarmellose sodium and calcium, examples of the lubricant include magnesium stearate, examples of the colorant include titanium oxide, and examples of of coating agent include hydroxypropyl methyl cellulose.

The solid formulation such as a tablet, capsule, granule or powder may normally contain any amount of the salts of the compound (I) and its crystals, as long as it exerts the efficacy to the point of being applicable as a medicine.

To produce an injection (for intravenous administration, for intramuscular administration, for subcutaneous administration, for intraperitoneal administration, and so on), a pH regulator, a buffering agent, a suspending agent, a solubility agent, an antioxidant, a preservative (antiseptic) and/or an agent isotonic to the salts of the compound (I) and its crystals, and an injection can be produced by a conventional method. The preparations can be lyophilized to transform them into extemporaneous dissolution type lyophilized preparations.

As the pH regulator and buffering agent, for example, an organic acid or an inorganic acid and/or a salt thereof can be used. As a suspending agent, hydroxypropyl cellulose can be used. As the solubilizing agent, polysorbate 80 can be used. As the antioxidant, α-tocopherol can be used. As a preservative, methyl para-hydroxybenzoate, ethyl para-hydroxybenzoate can be used. As isotonic agent, glucose can be used.

The injection formation usually contains any amount of the salts of the compound (I) and its crystals, as long as it exerts the efficacy to the point of being applicable as a medicine.

To produce the external formulation, a base raw material is added to the salts of the compound (I) and its crystals, and if necessary, for example, the preservative, stabilizer, pH regulator, antioxidant and/or are added. or the colorant described above, and for example, a skin preparation (ointment, patch and the like), eye drops, nasal drops and/or suppository can be produced by conventional methods.

As base raw materials to be used, various raw materials are usually used, for example, for medicines, quasi-drugs and cosmetics can be used. Specific examples of these include raw materials such as animal and vegetable oils, mineral oils, ester-type oils, waxes, emulsifiers, higher alcohols, fatty acids, silicone oils, surfactants, phospholipids, alcohols, polyhydric alcohols, water-soluble polymers, clay minerals. and purified water.

The external preparation may normally contain any amount of salts of the compound (I) and its crystals, as long as it exerts efficacy in the sense of being applicable as a medicine.

A dose of the salts of compound (I) and their crystals depends on the level of severity of the symptoms, the patient's age, sex and weight, the form of administration and the type of salt and/or the specific type of disease, and is not specially limited unless it exceeds the maximum dose of the medicine that can be given without causing an unacceptable adverse reaction, and in an adult patient, it is administered, once a day or divided into several times a day, at a dose to oral administration, generally about 30 pg to 10 g, specifically 100 pg to 5 g, and more specifically 100 micrograms to 1 g, or a dose for administration by injection, generally about 30 pg to 1 g , specifically from 100 pg to 500 mg, and more specifically from 100 pg to 300 mg.

Example

The compounds according to the present invention can be produced by the methods described in the Production Examples and Examples described below, for example.

In X-ray powder diffractometry of the crystals produced in the following Examples and Reference Examples, the resulting crystals were placed in a sample stage of an X-ray powder diffractometer under the following conditions. Figures 1-3 and 6-15 show the results.

Measurement conditions

Sample holder: aluminum

Objective: copper

Detector: scintillation counter

Tube voltage: 50 kV

Tube current: 300mA

Slit: 0.5mm DS (2mm height limiting slit), Open SS, Open RS

Sweep speed: 10°/min

Sampling interval: 0.02°

Sweep range: 5 to 35°

The 13 C solid state NMR spectra of the crystals were measured under the following conditions. Figure 4 shows the result.

Measurement conditions

Device used: AVANCE400 (from Bruker Corporation)

Measurement temperature: room temperature (22 °C)

Reference material: glycine (external reference: 176.03 ppm)

Measured core: 13C (100.6248425 MHz)

Pulse repetition time: 3 seconds

Pulse mode: TOSS measurement

In the Production Examples, silica gel 60 (Kanto Chemicals) or Silica Gel Presep (WAKO) was used as the purification silica gel for silica gel column chromatography unless otherwise stated. In addition, NH silica gel (Fuji Silysia Chemical LTD.) or Hi-Flash Column Amino (YAMAZENE CORPORATION) was used as the purification silica gel used for NH silica gel column chromatography.

Varian Mercury 400, Varian Mercury Plus 400, Varian INOVA 500, or Avance 600 MHz (Bruker) was used for the measurement of proton nuclear magnetic resonance spectra, and proton nuclear magnetic resonance spectra were measured at 400 MHz at minus that the contrary be asserted. Shifts of proton nuclear magnetic resonance spectra are recorded in the unit of o (ppm) relative to tetramethylsilane and coupling constants are recorded in the unit of Hertz (Hz). Abbreviations for spacing patterns are as follows: s: singlet; d: doublet; t: triplet; m: multiplet; and brs: broad singlet.

In Production Examples, Examples and Reference Examples, commercially available products were appropriately used as commercially available compounds.

[Production Example 1-1] N-(4-chloropyridin-2-yl)acetamide

Commercially available 2-amino-4-chloropyridine (50 g, 389 mmol) was dissolved in acetic anhydride (500 mL), triethylamine (271 mL, 1.94 mol) was added at 20°C, and stirred. the resulting mixture at 60°C for 12 hours. The mixture was cooled to room temperature and then the solvent was evaporated. The residue was purified with silica gel column chromatography (n-heptane:ethyl acetate = 4 : 1 to 1 : 1), and then the target fraction was concentrated in vacuo to obtain the title compound (66 g, 99% ).

1H-NMR Spectrum (CDCla) 5 (ppm): 2.21 (3H, s), 7.05 (1H, dd, J = 5.4, 1.9 Hz), 8.15 (1H, d, J = 5.4 Hz), 8.30 (2H, s a ).

[Production example 1-2] Phenyl methylcarbamate

A mixture of commercially available methylamine hydrochloride (50 g, 0.74 mol), pyridine (124 mL, 1.53 mol), and N,N-dimethylformamide (500 mL) was stirred at 5°C, and sodium chlorocarbonate was added dropwise. commercially available phenyl (94 mL, 0.75 mol) for 2 hours. After complete addition, the mixture was stirred under nitrogen at room temperature for 16 hours. The reaction mixture was added to ice water (2 L) and extracted with ethyl acetate (1.5 L) twice. The organic phase was washed with water (1 L) and saturated brine (300 mL). The organic phase was dried over anhydrous magnesium sulfate, and then the solvent was evaporated. n-Heptane and ethyl acetate were added to the concentrator residue and the precipitate was collected by filtration and washed with n-heptane and methyl t-butyl ether to obtain the title compound (74.2 g, 66%).

1H-NMR Spectrum (CDCh) 5 (ppm): 2.90 (3H, d, J = 4.9 Hz), 4.95 (1H, s a), 7.08-7.16 (2H, m), 7.16-7.24 (1H, m), 7.31 7.41 (2H, m)

[Production Example 1-3] 1-(4-phenylpiperidin-1-yl)ethanone

A mixture of commercially available 4-phenylpiperidine (10 g, 62 mmol), pyridine (5.7 mL, 70.5 mmol), and tetrahydrofuran (80 mL) was stirred at 0°C and acetyl chloride mixture (5 mL, 70.3 mmol) and tetrahydrofuran (20 mL) for 10 minutes. The mixture was stirred under a nitrogen atmosphere at 25°C for 14 hours. Acetate (100 ml) and water (100 ml) were added to the reaction liquid for separation. The aqueous phase was extracted with ethyl acetate (100 mL), then the organic phases were combined, and the resulting portion was washed with aqueous sodium bicarbonate solution (100 mL), water (100 mL), and then brine. saturated (50 ml). The organic layer was dried over anhydrous magnesium sulfate, then the solvent was evaporated to give the title compound (12.3 g, 98%).

1H-NMR Spectrum (CDCla) 5 (ppm): 1.52-1.78 (2H, m), 1.81-1.99 (2H, m), 2.14 (3H, s), 2.63 (1H, td, J = 12.9, 2.7 Hz) , 2.74 (1H, tt, J = 12.1, 3.7 Hz), 3.17 (1H, td, J = 13.2, 2.6 Hz), 3.84-4.02 (1H, m), 4.69-4.89 (1H, m), 7.08-7.43 (5H, m).

[Production example 1-4] 4-(1-acetylpiperidin-4-yl)benzoic acid

A mixture of aluminum(III) chloride (26 g, 195 mmol) and dichloromethane (200 mL) was stirred at 0°C, and oxalyl chloride (20 mL, 228 mmol) was added over 10 minutes. A mixture of 1-(4-phenylpiperidin-1-yl)ethanone described in Production Example 1-3 (12.3 g, 60.5 mmol) and dichloromethane (50 mL) was then added over 30 minutes. The mixture was stirred under a nitrogen atmosphere at 25°C for 14 hours. The reaction liquid was poured onto ice, and ethyl acetate (1 L) and water (1 L) were added for separation. The aqueous phase was extracted with ethyl acetate (1 L) twice, then the organic phase was washed with water (1 L) twice and then with a saturated saline solution (500 mL). The organic phase was dried over anhydrous magnesium sulfate, and then the solvent was evaporated. Ethyl acetate was added to the concentrator residue and the product was collected by filtration and washed with ethyl acetate to give the title compound (9.09 g, 61%).

1H-NMR Spectrum (CDCla) 5 (ppm): 1.49-1.82 (2H, m), 1.92 (2H, t, J = 13.2 Hz), 2.15 (3H, s), 2.65 (1H, t, J = 11.7 Hz ), 2.75-2.94 (1H, m), 3.08-3.30 (1H, m), 3.97 (1H, d, J = 13.2 Hz), 4.82 (1H, d, J = 12.8 Hz), 7.30 (2H, d, J = 8.4 Hz), 8.05 (2H, d, J = 8.1 Hz).

[Production example 1-5] 4-(piperidin-4-yl)benzoic acid hydrochloride

A mixture of 4-(1-acetylpiperidin-4-yl)benzoic acid in Production Example 1-4 (4.50 g, 18.2 mmol) and 5 M hydrochloric acid (50 mL, 250 mmol) was stirred under nitrogen at 140°C for 18 hours. The mixture was cooled to room temperature, then the product was collected by filtration and washed with water to obtain the title compound (3.77 g, 86%).

1H-NMR Spectrum (DMSO-ds) 5 (ppm): 1.60-2.15 (4H, m), 2.76-3.16 (3H, m), 3.27-3.45 (2H, m), 7.36 (2H, d, J = 8.1 Hz), 7.92 (2H, d, J = 8.1 Hz), 8.65-9.04 (2H, m), 12.89 (1H, sa).

[Production example 1-6] 4-(1-(tert-butoxycarbonyl)piperidin-4-yl)benzoic acid

A mixture of 4-(piperidin-4-yl)benzoic acid hydrochloride described in Production Example 1-5 (2.00 g, 8.27 mmol), 1 M sodium hydroxide solution (25 mL, 25 mmol) was stirred. and acetone (50 mL) at 25°C, and a solution of di-tert-butyl dicarbonate (1.9 g, 8.71 mmol) in acetone (25 mL) was added dropwise over 10 minutes. The mixture was stirred under a nitrogen atmosphere at 25°C for 18 hours. 1M Hydrochloric acid (17 ml) was added under cooling at 0°C. The mixture was extracted with ethyl acetate (100 ml) twice. The organic phase was washed with saturated brine (50 mL). The organic phase was dried over anhydrous magnesium sulfate and then concentrated in vacuo. n-Heptane and methyl and tert-butyl ether were added to the concentrated residue and the product was collected by filtration and washed with n-heptane to obtain the title compound (2.30 g, 91%).

1H-NMR Spectrum (CDCls) 5 (ppm): 1.49 (9H, s), 1.57-1.76 (2H, m), 1.84 (2H, d, J = 13.5 Hz), 2.62-2.97 (3H, m), 4.27 (2H, s a), 7.28-7.36 (2H, m), 7.98-8.10 (2H, m).

[Production example 1-7] 3-hydroxy-4-(2-methoxyethoxy)benzaldehyde

Commercially available 3,4-dihydroxybenzaldehyde (39.3 g, 285 mmol) and sodium carbonate (45.2 g, 427 mmol) were dissolved in N,N-dimethylformamide (400 mL), then 2-bromoethyl methyl ether (26.7 mL, 285 mmol) commercially available under nitrogen atmosphere at room temperature, and the mixture was stirred for 5 days. The mixture was cooled to 0°C, and then 2M hydrochloric acid, ethyl acetate and water were added for separation. The aqueous phase was extracted with ethyl acetate, then the organic phase was washed with saturated brine and dried over anhydrous magnesium sulfate, and then filtered. The solvent was evaporated, dichloromethane was added, the precipitate was separated by filtration, and then the resulting filtrate was purified with silica gel column chromatography (n-heptane:ethyl acetate = 17:3 to 1:1 ). The target fraction was concentrated in vacuo to obtain the title compound (12.9 g, 23%).

1H-NMR Spectrum (CDCla) 5 (ppm): 3.47 (3H, s), 3.76-3.80 (2H, m), 4.25-4.29 (2H, m), 6.40 (1H, s a), 7.01 (1H, d, J = 8.4 Hz), 7.41 (1H, dd, J = 8.2, 2.0 Hz), 7.45 (1H, d, J = 1.8 Hz), 9.85 (1H, s).

[Production Example 1-8] 3-(benzyloxy)-4-(2-methoxyethoxy)benzaldehyde

Potassium carbonate (11.8 g, 85.7 mmol) and benzyl chloride (10 mL, 86.9 mmol) were added to the liquid mixture of 3-hydroxy-4-(2-methoxyethoxy)benzaldehyde described in Production Example 1-7 ( 12.9 g, 65.9 mmol) in ethanol (130 mL) under nitrogen atmosphere at room temperature, and the mixture was heated under reflux at 90°C for 2 hours. The mixture was cooled to 0°C, and then 2M hydrochloric acid, ethyl acetate and water were added for separation. The organic phase was washed with a saturated saline solution, dried over anhydrous magnesium sulfate, and then filtered. The solvent was evaporated, and the resulting residue was purified with silica gel column chromatography (nheptane:ethyl acetate = 9:1 to 1:1). The target fraction was concentrated in vacuo to obtain the title compound (17.6 g, 93%).

1H-NMR Spectrum (CDCla) 5 (ppm): 3.46 (3H, s), 3.79-3.85 (2H, m), 4.24-4.30 (2H, m), 5.18 (2H, s), 7.03 (1H, d, J = 8.1 Hz), 7.29-7.35 (1H, m), 7.35-7.41 (2H, m), 7.43-7.50 (4H, m), 9.82 (1H, s).

[Production Example 1-9] (E)-2-(benzyloxy)-1-(2-methoxyethoxy)-4 -(2-nitrovinyl)benzene

3-(Benzyloxy)-4-(2-methoxyethoxy)benzaldehyde described in Production Example 1-8 (17.6 g, 61.5 mmol) was dissolved in acetic acid (49.3 mmol), then ammonium acetate (5.69 g, 73.8 mmol) and nitromethane (8.32 mL, 154 mmol) under nitrogen atmosphere at room temperature, and the mixture was heated under reflux at 130°C for 2 hours. The mixture was cooled to room temperature, and then the precipitate was collected by filtration and washed with ethanol to obtain the title compound quantitatively.

1H-NMR Spectrum (CDCls) 5 (ppm): 3.46 (3H, s), 3.78-3.84 (2H, m), 4.21-4.27 (2H, m), 5.16 (2H, s), 6.97 (1H, d, J = 8.4 Hz), 7.06 (1H, d, J = 1.8 Hz), 7.16 (1H, dd, J = 8.4, 2.2 Hz), 7.30-7.48 (6H, m), 7.91 (1H, d, J = 13.5 Hz).

[Production example 1-10] 6-(2-methoxyethoxy)-1H-indole-5-ol

Nitric acid 69% (15 mL, 233 mmol) was added to a mixture of (E)-2-(benzyloxy)-1-(2-methoxyethoxy)-4-(2-nitrovinyl)benzene described in the Production Example 1-9 (20.2 g, 61.5 mmol) and acetic acid (120 ml) at 25°C, and the mixture was stirred at room temperature for 6 hours. The reaction mixture was poured onto ice, and the precipitate was collected by filtration, and then washed with water to obtain a crude product (23.0 g).

The crude product (23.0 g) was suspended in methanol (500 ml), then palladium-carbon 10% (water content, 50%) (8 g) was added at room temperature, and the mixture was stirred under hydrogen atmosphere. for 6 hours. The catalyst was filtered off with celite, the filtrate was concentrated in vacuo, and the resulting part was purified with silica gel column chromatography (n-heptane:ethyl acetate = 2:1 to 1:1). The target fraction was concentrated in vacuo to obtain the title compound (3.94 g, 31%).

1H-NMR Spectrum (CDCla) 5 (ppm): 3.48 (3H, s), 3.69-3.78 (2H, m), 4.16-4.23 (2H, m), 6.24 (1H, s), 6.41 (1H, ddd, J = 3.1,2.1,0.8 Hz), 6.97 (1H, s), 7.10 (1H, dd, J = 3.2, 2.5 Hz), 7.15 (1H, s), 7.94 (1H, s a).

[Production Example 1-11] N-(4-((6-(2-methoxyethoxy)-1H-indol-5-yl)oxy)pyridin-2-yl)acetamide

6-(2-Methoxyethoxy¡)-1H-indol-5-ol described in Production Example 1-10 (3.94 g, 19.0 mmol) and N-(4-chloropyridin-2-yl)acetamide described in Production Example 1-1 (3.25 g, 19.0 mmol) in dimethylsulfoxide (25 mL), then 97% potassium tert-butoxide (2.20 g, 19.0 mmol) was added at room temperature, and the mixture was warmed and stirred. at 150°C for 13 hours. Water and ethyl acetate were added to the reaction liquid at room temperature for separation. The aqueous phase was extracted with ethyl acetate three times and the combined organic phase was washed with water. The organic phase was dried over anhydrous sodium sulfate. The drying agent was filtered off, and the filtrate was concentrated in vacuo, and the resulting part was purified with silica gel column chromatography NH(n-heptane:ethyl acetate=2:3 to 0:1-ethyl acetate:methanol ) from 49:1 to 9:1). The target fraction was concentrated in vacuo to obtain the title compound (3.45 g, 53%).

1H-NMR Spectrum (500MHz, CDCls) 5 (ppm): 2.13 (3H, s), 3.27 (3H, s), 3.54-3.58 (2H, m), 4.07-4.11 (2H, m), 6.46 6.50 (1H , m), 6.54 (1H, dd, J = 5.8, 1.9 Hz), 7.05 (1H, s), 7.14-7.17 (1H, m), 7.36 (1H, s), 7.75 (1H, sa), 8.02 ( 1H, d, J = 5.8 Hz), 8.10 (1H, brs), 8.19 (1H, brs).

[Production Example 1-12] 4-((6-(2-methoxyethoxy)-1 H-indol-5-yl)oxy)pyridin-2-amine

N-(4-((6-(2-methoxyethoxy)-1H-indol-5-yl)oxy)pyridin-2-yl)acetamide described in Production Example 1-11 (3.45 g, 10.1 mmol) was dissolved in methanol (50 mL), 2M sodium hydroxide solution (50 mL) was added at room temperature, and the mixture was heated at 70°C for 3 hours. Water and ethyl acetate were added to the reaction mixture for separation. The aqueous phase was extracted with ethyl acetate three times, and the combined organic phase was dried over anhydrous sodium sulfate. The drying agent was filtered off, and the filtrate was concentrated in vacuo, and the resulting part was purified with silica gel column chromatography NH(n-heptane:ethyl acetate=3:7 to 0:1-ethyl acetate: methanol = from 49:1 to 24:1). The target fraction and the mixed fraction were concentrated in vacuo separately from each other, the mixed fraction was purified again with silica gel column chromatography (ethyl acetate:methanol = 1:0 to 9:1) and then the resulting part was combined with the objective fraction described above to obtain the title compound (2.60 g, 86%).

1H-NMR Spectrum (500MHz, CDCh) or (ppm): 3.31 (3H, s), 3.58-3.63 (2H, m), 4.08-4.11 (2H, m), 4.28 (2H, s a), 5.90 (1 H , d, J = 2.4 Hz), 6.29 (1H, dd, J = 6.1,2.2 Hz), 6.44-6.52 (1H, m), 7.06 (1H, s), 7.15-7.20 (1H, m), 7.34 (1H, s), 7.88 (1H, d, J = 5.8 Hz), 8.22 (1H, s a).

[Production Example 1-13] 5-[(2-aminopyridin-4-yl)oxy]-6-(2-methoxyethoxy)-N-methyl-1H-indole-1-carboxamide

4-((6-(2-Methoxyethoxy)-1H-indol-5-yl)ox)pyridin-2-amine described in Production Example 1-12 (2.60 g, 8.67 mmol) was dissolved in N,N- dimethylformamide (50 mL), then oily sodium hydride 50-72% (499 mg) was added under nitrogen at room temperature. Phenyl methylcarbamate from Production Example 1-2 (1.97 g, 13.0 mmol) was added, and the mixture was stirred at room temperature for 1 hour. The reaction mixture was cooled to 0°C and ethyl acetate and water were added for separation. The aqueous phase was extracted with ethyl acetate twice, sodium chloride was added to the aqueous phase, and the resulting part was extracted with ethyl acetate three times. The combined organic phase was dried over anhydrous sodium sulfate. The drying agent was filtered off, and the filtrate was concentrated in vacuo, and then purified with silica gel column chromatography NH(n-heptane:ethyl acetate=1:4 to 0:1-ethyl acetate:methanol = from 49:1 to 24:1). The target fraction was concentrated in vacuo, and ethyl acetate was added and the precipitate was collected by filtration and washed to obtain the title compound (2.23 g, 72%).

1H-NMR Spectrum (500MHz, CDCh) 5 (ppm): 3.06 (3H, d, J = 4.9 Hz), 3.29 (3H, s), 3.59-3.63 (2H, m), 4.14-4.17 (2H, m) , 4.30 (2H, s a), 5.52-5.59 (1 H, m), 5.89 (1 H, d, J = 2.4 Hz), 6.27 (1H, dd, J = 5.8, 1.9 Hz), 6.55 (1 H, d, J = 3.9 Hz), 7.27-7.29 (2H, m), 7.89 (1H, d, J = 5.9 Hz), 7.99 (1H, s).

[Production Example 1-14] 6-(2-methoxyethoxy)-N-methyl-5-{[2-({[4-(piperidin-4-yl)phenyl]carbonyl}amino)pyridin-4-yl] oxy}-1H-indole-1-carboxamide

Benzotriazole (609 mg, 5.11 mmol) was dissolved in dichloromethane (25 ml), thionyl chloride (373 gl, 5.11 mmol) was added under nitrogen at room temperature and the mixture was stirred for 5 minutes. 4-(1-(tert-Butoxycarbonyl)piperidin-4-yl)benzoic acid described in Production Example 1-6 (1.3 g, 4.26 mmol) was added to the reaction mixture at room temperature, and the mixture was stirred for 30 minutes. The reaction mixture was filtered through a glass fiber filter completely covered with anhydrous sodium sulfate, and the resulting part was washed with dichloromethane, the filtrate was added to a mixture of 5-[(2-aminopyridin-4-yl )oxy]-6-(2-methoxyethoxy)-N-methyl-1H-indole-1-carboxamide described in Production Example 1-13 (0.95 g, 2.67 mmol), triethylamine (1.86 mL, 13.3 mmol), and 4-dimethylaminopyridine (16 mg, 0.133 mmol) in N,N-dimethylformamide (3 mL) and dichloromethane (20 mL) at 0°C for 5 min, and the mixture was washed with dichloromethane (10 mL), then stirred at the same temperature for 5 min. The mixture was stirred at room temperature for 2 hours, then 40% methylamine aqueous solution (2.3 mL, 26.7 mmol) was added, and then the mixture was stirred at room temperature for 1.5 hours. A saturated aqueous solution of sodium bicarbonate was added to the reaction mixture for separation, and the aqueous phase was extracted with ethyl acetate three times. The combined organic phase was dried over anhydrous sodium sulfate. The drying agent was filtered off, then the filtrate was concentrated in vacuo, and the resulting part was purified with silica gel column chromatography (n-heptane:ethyl acetate=1:1 to 0:1-ethyl acetate :methanol = 49:1 to 23:2) to obtain the crude product (1.11 g).

The crude product (1.11 g) was dissolved in dichloromethane (50 ml) and trifluoroacetic acid (5.0 ml) was added at room temperature. The mixture was stirred at room temperature for 30 minutes, then the resulting part was concentrated in vacuo, and then the residue was dissolved in dichloromethane and triethylamine, and the resulting part was concentrated in vacuo. The residue was purified with NH silica gel column chromatography (ethyl acetate:methanol = 1:0 to 22:3) to obtain the title compound (829 mg, 57%).

1H-NMR Spectrum (500MHz, CDCh) 5 (ppm): 1.59-1.69 (2H, m), 1.83 (2H, d, J = 14.1 Hz), 2.68 (1H, tt, J = 12.0, 3.6 Hz), 2.75 (2H, td, J = 12.2, 2.4 Hz), 3.04 (3H, d, J = 4.9 Hz), 3.17-3.23 (2H, m), 3.26 (3H, s), 3.55-3.61 (2H, m), 4.15 4.21 (2H, m), 5.57-5.65 (1 H, m), 6.53 (1H, d, J = 3.4 Hz), 6.62 (1H, dd, J = 5.8, 2.4 Hz), 7.25 (1 H, d , J = 3.9 Hz), 7.30 7.34 (3H, m), 7.77-7.82 (2H, m), 7.91 (1 H, d, J = 2.4 Hz), 8.02 (1H, s), 8.10 (1 H, d , J = 5.9 Hz), 8.50 (1H, s a).

[Production Example 1-15] 5-({2-[({4-[1-(2-hydroxyethyl)piperidin-4-yl]phenyl}carbonyl)amino]pyridin-4-yl}oxy)-6- (2-methoxyethoxy)-N-methyl-1H-indole-1-carboxamide

Sodium triacetoxyborohydride (114 mg, 0.54 mmol) and commercially available 2-hydroxyacetaldehyde (34.4 mg, 0.57 mmol) were added to a mixture of 6-(2-methoxyethoxy)-N-methyl-5-{[2-({[ 4-(piperidin-4-yl)phenyl]carbonyl}amino)pyridin-4-yl]oxy}-1 H-indole-1-carboxamide described in Production Example 1-14 (100 mg, 0.18 mmol) and tetrahydrofuran (4 ml) at room temperature, and the mixture was stirred at room temperature for 2 hours. A saturated aqueous solution of sodium bicarbonate and ethyl acetate were added to the reaction mixture for separation. The aqueous phase was extracted with ethyl acetate, and the combined organic phase was washed with a saturated saline solution, then dried over anhydrous sodium sulfate, and then filtered. The solvent was evaporated, and the resulting residue was purified with NH silica gel column chromatography (ethyl acetate:methanol = 1:0 to 97:3-9:1). The target fraction was concentrated in vacuo, then the precipitate was collected by filtration and washed with a liquid mixture of diethyl ether and n-hexane to obtain the title compound (90 mg, 83%). Typical powder X-ray diffraction angles of the obtained compound (free form of compound (I) (Free Form A)) are shown below. (20 ± 0.2°): 10.4°, 10.9°, 11.4°, 13.5°, 16.1°, 19.7°, 20.4°, 21.5°, 23.3° and 24.3°.

1H-NMR Spectrum (CDCh) 5 (ppm): 1.70-1.92 (4H, m), 2.15-2.24 (2H, m), 2.53-2.65 (3H, m), 3.01-3.09 (5H, m), 3.26 ( 3H, s), 3.56-3.60 (2H, m), 3.64 (2H, t, J = 5.2 Hz), 4.15-4.20 (2H, m), 5.49-5.54 (1H, m), 6.55 (1H, d , J = 3.7 Hz), 6.61 (1H, dd, J = 5.8, 2.3 Hz), 7.24-7.28 (1H, m), 7.30-7.35 (3H, m), 7.81 (2H, d, J = 8.2 Hz), 7.91 (1H, d, J = 2.4 Hz), 8.01 (1 H, s), 8.10 (1H, d, J = 5.9 Hz), 8.50 (1 H, s a).

[Example 1]

Preparation of a crystal of 5-({2-[({4-[1-(2-hydroxyethyl)piperidin-4-yl]phenyl}carbonyl)amino]pyridin-4-yl}oxy)- succinate salt 6-(2-methoxyethoxy)-N-methyl-1 H-indole-1-carboxamide (other name: 5-({2-[({4-[1-(2-hydroxyethyl)piperidin-4-yl)phenyl }carbonyl)amino]pyridin-4-yl}oxy)-6-(2-methoxyethoxy)-N-methyl-1 H-indole-1-carboxamide=butanedioate (2:3))

2.93 g of 5-({2-[({4-[1-(2-hydroxyethyl)piperidin-4-yl]phenyl}carbonyl)amino]pyridin-4-yl}oxy)-6-(2- methoxyethoxy)-N-methyl-1H-indole-1-carboxamide described in Production Example 1-15, in a recovery flask, 60 ml of ethanol was added, and the mixture was heated and stirred at 70°C. in an oil bath for dissolution. Succinic acid (1.23 g) was added, then the oil bath was quenched and gradually cooled. The mixture was stirred at room temperature for 2 hours, and then stirred at 5°C for 1 hour. The solid was collected by filtration to obtain the title compound (3.70 g).

1H-NMR Spectrum (600MHz, CDaOD) 5 (ppm): 1.96-2.10 (4H, m), 2.52 (6H, s), 2.93 (1H, m), 2.96 (3H, s), 3.01 (2H, m) , 3.16 (2H, t, J=5.4 Hz), 3.22 (3H, s), 3.56 (2H, t, J=4.7 Hz), 3.61 (2H, m), 3.87 (2H, t, J=5.4 Hz) , 4.14 (2H, t, J=4.6 Hz), 6.61 (1H, d, J=3.6 Hz), 6.68 (1H, dd, J=5.8, 2.3 Hz), 7.37 (1H, s), 7.42 (2H, d, J=8.3 Hz), 7.58 (1H, d, J=3.6 Hz), 7.73 (1H, d, J=2.2 Hz), 7.88 (2H, d, J=8.3 Hz), 8.08 (1H, s) , 8.15 (1H, d, J=5.8 Hz).

13C-NMR(100 MHz, solid state) 5 (ppm): 27.1,28.3, 29.7, 34.8, 38.0, 41.3, 54.0, 57.3, 59.7, 60.9, 72.1,72.5, 103.3, 104.2, 108.5, 116.9, 128.9 , 134.5, 136.7, 140.7, 149.4, 151.3, 155.1, 169.5, 170.1, 175.6, 179.9, 183.7.

[Example 2]

Preparation of a crystal of 0.5 5-({2-[({4-[1-(2-hydroxyethyl)piperidin-4-yl]phenyl}carbonyl)amino]pyridin-4-yl}oxy)- succinate salt 6-(2-methoxyethoxy)-N-methyl-1H-indole-1-carboxamide(a)

117 mg of 5-({2-[({4-[1-(2-hydroxyethyl)piperidin-4-yl]phenyl}carbonyl)amino]pyridin-4-yl}oxy)-6-(2- methoxyethoxy)-N-methyl-1H-indole-1-carboxamide described in Production Example 1-15 and succinic acid (11.8 mg) to a 30 mL recovery flask, 2 mL isopropanol/water solution was added (8/2, v/v), ultrasonic irradiation was carried out, and the mixture was stirred at room temperature for 2-3 hours. The solid was collected by filtration to obtain the title compound (77.5 mg).

1H-NMR Spectrum (600MHz, CDaOD) 5 (ppm): 1.86-2.00 (4H, m), 2.51 (2H, s), 2.62 (2H, m), 2.79 (1H, m), 2.87 (2H, t, J=5.5 Hz), 2.96 (3H, s), 3.22 (3H, s), 3.36 (2H, d, J=11.8 Hz), 3.56 (2H, t, J=4.6 Hz), 3.79 (2H, t, J=5.7 Hz), 4.15 (2H, t, J=4.6 Hz), 6.61 (1H, d, J=3.6 Hz), 6.68 (1H, dd, J=5.7, 2.1 Hz), 7.37 (1H, s) , 7.40 (2H, d, J=8.2 Hz), 7.58 (1H, d, J=3.6 Hz), 7.73 (1H, d, J=2.0 Hz), 7.86 (2H, d, J=8.3 Hz), 8.08 (1H, s), 8.14 (1H, d, J=5.8 Hz).

[Example 3]

Preparation of a crystal of 5-({2-[({4-[1-(2-hydroxyethyl)piperidin-4-yl]phenyl}carbonyl)amino]pyridin-4-yl}oxy)-6 maleate salt -(2-methoxyethoxy)-N-methyl-1H-indole-1-carboxamide

Maleic acid (24.1 mg) and 2 mL of acetone were added to 101 mg of 5-({2-[({4-[1-(2-hydroxyethyl)piperidin-4-yl]phenyl}carbonyl)amino]pyridin- 4-yl}oxy)-6-(2-methoxyethoxy)-N-methyl-1H-indole-1-carboxamide described in Production Example 1-15, and the mixture was stirred at room temperature. The solid was collected by filtration to obtain the title compound (113mg).

[Example 4]

Preparation of a crystal of 0.5 5-({2-[({4-[1-(2-hydroxyethyl)piperidin-4-yl]phenyl}carbonyl)amino]pyridin-4-yl}oxy)- succinate salt 6-(2-methoxyethoxy)-N-methyl-1H-indole-1-carboxamide (a)

550 mg of 5-({2-[({4-[1-(2-hydroxyethyl)piperidin-4-yl]phenyl}carbonyl)amino]pyridin-4-yl}oxy)-6-(2- methoxyethoxy)-N-methyl-1H-indole-1-carboxamide, succinic acid (55.3 mg) and water (5.5 ml) were added to a test tube, and ultrasonic irradiation was carried out, and then the mixture was stirred at room temperature. environment at night. The solid was filtered overnight. The solid was ground in an agate mortar, and stored under the condition of 40°C/75%RH for about 1.5 hours, and then the title compound (620 mg) was obtained.

1H-NMR Spectrum (CD ³ OD) 5 (ppm): 1.86-2.00 (4H, m), 2.51 (2H, s), 2.62 (2H, m), 2.79 (1H, m), 2.87 (2H, rt, J=5.5 Hz), 2.96 (3H, s), 3.22 (3H, s), 3.36 (2H, da, J=11.8 Hz), 3.56 (2H, ta, J=4.6 Hz), 3.79 (2H, t, J=5.7 Hz), 4.15 (2H, ta, J=4.6 Hz), 6.61 (1H, d, J=3.6 Hz), 6.68 (1H, dd, J=5.7, 2.1 Hz), 7.37 (1H, s) , 7.40 (2H, d, J=8.2 Hz), 7.58 (1H, d, J=3.6 Hz), 7.73 (1H, d, J=2.0 Hz), 7.86 (2H, d, J=8.3 Hz), 8.08 (1H, s), 8.14 (1H, d, J=5.8 Hz)

[Example 5]

Preparation of a crystal of 0.5 5-({2-[({4-[1-(2-hydroxyethyl)piperidin-4-yl]phenyl}carbonyl)amino]pyridin-4-yl}oxy)- succinate salt 6-(2-methoxyethoxy)-N-methyl-1 H-indole-1 -carboxamide 0.5 (p)

The sample obtained in Example 4 was dried under reduced pressure for 3 days to obtain the title compound.

1H-NMR Spectrum (CD ³ OD) 5 (ppm): 1.87-2.00 (4H, m), 2.51 (2H, s), 2.65 (2H, m), 2.79 (1H, m), 2.89 (2H, rt, J=5.6 Hz), 2.95 (3H, s), 3.22 (3H, s), 3.38 (2H, da, J=12.1 Hz), 3.56 (2H, m), 3.80 (2H, t, J=5.6 Hz) , 4.14 (2H, m), 6.60 (1H, d, J=3.7 Hz), 6.67 (1H, dd, J=5.8, 2.3 Hz), 7.37 (1H, s), 7.40 (2H, d, J=8.4 Hz), 7.57 (1H, d, J=3.7 Hz), 7.73 (1H, d, J=2.3 Hz), 7.86 (2H, d, J=8.4 Hz), 8.08 (1H, s), 8.14 (1H, d, J=5.8Hz)

[Example 6]

Preparation of 1.5 5-({2-[({4-[1-(2-hydroxyethyl)piperidin-4-yl]phenyl}carbonyl)amino]pyridin-4-yl}oxy)-6-( 2-methoxyethoxy)-N-methyl-1H-indole-1-carboxamide

251 mg of 1.5 5-({2-[({4-[1-(2-hydroxyethyl)piperidin-4-yl]phenyl}carbonyl)amino]pyridin-4-yl}oxy)- succinate salt were dissolved 6-(2-methoxyethoxy)-N-methyl-1H-indole-1-carboxamide in 25 mL of 50% tert-butyl alcohol aqueous solution. 3 ml of sample solution was added to a test tube, and the sample solution in ethanol was frozen with dry ice. Solvent was removed on lyophilizer to obtain the title compound (30.8 mg).

1H-NMR Spectrum (CD ³ OD) 5 (ppm): 1.96-2.11 (4H, m), 2.53 (6H, s), 2.93 (1H, m), 2.96 (3H, s), 3.00 (2H, m) , 3.15 (2H, t, J=5.4 Hz), 3.22 (3H, s), 3.56 (2H, m), 3.60 (2H, da, J=12.4 Hz), 3.87 (2H, t, J=5.5 Hz) , 4.15 (2H, ta, J=4.6 Hz), 6.61 (1H, d, J=3.7 Hz), 6.68 (1H, da, J=3.8 Hz), 7.37 (1H, s), 7.42 (2H, d, J=8.3 Hz), 7.58 (1H, d, J=3.8 Hz), 7.73 (1H, sa), 7.88 (2H, d, J=8.3 Hz), 8.08 (1H, s), 8.15 (1H, da, J=5.0Hz)

[Reference Example 1]

Preparation of a crystal of 5-({2-[({4-[1-(2-hydroxyethyl)piperidin-4-yl]phenyl}carbonyl)amino]pyridin-4-yl}oxy)-6-(2- methoxyethoxy)-N-methyl-1H-indole-1-carboxamide (Free Form B)

93.2 mg of 5-({2-[({4-[1-(2-hydroxyethyl)piperidin-4-yl]phenyl}carbonyl)amino]pyridin-4-yl}oxy)-6-(2- methoxyethoxy)-N-methyl-1H-indole-1-carboxamide described in Production Example 1-15 in a test tube, then 2.99 ml of isopropanol and 264 μl of water were added. The mixture was heated to 70-100°C in an oil bath for dissolution. The resulting solution was stirred at -5°C in a thermostatically controlled bath for 16 hours. The solid precipitate was collected by filtration and dried under reduced pressure overnight to obtain the title compound. The powder X-ray diffraction angles of the obtained compound are shown below. (20 ± 0.2°): 7.8°, 10.8°, 13.1°, 14.2°, 17.8°, 21.5°, 21.7°, 23.4°, 24.5° and 29.0°.

[Reference Example 2]

Preparation of a crystal of 5-({2-[({4-[1-(2-hydroxyethyl)piperidin-4-yl]phenyl}carbonyl)amino]pyridin-4-yl}oxy)-6-(2- methoxyethoxy)-N-methyl-1H-indole-1-carboxamide (Hydrate Free Form)

2 ml of isopropanol and 2 ml of water were added to 208 mg of 5-({2-[({4-[1-(2-hydroxyethyl)piperidin-4-yl]phenyl}carbonyl)amino]pyridin-4- yl}oxy)-6-(2-methoxyethoxy)-N-methyl-1H-indole-1-carboxamide described in Production Example 1-15. After carrying out ultrasonic irradiation in ice water, the mixture was stirred at 5°C for 3 days. The suspended solid was collected by filtration to obtain the title compound (106 mg). The powder X-ray diffraction angles of the obtained compound are shown below. (20 ± 0.2°): 8.8°, 9.6°, 15.2°, 16.3°, 20.0°, 20.8°, 21.4°, 22.0°, 23.8° and 27.1°.

[Reference Example 3]

Preparation of a crystal of 5-({2-[({4-[1-(2-hydroxyethyl)piperidin-4-yl]phenyl}carbonyl)amino]pyridin-4-yl}oxy)-6 mesylate salt -(2-methoxyethoxy)-N-methyl-1H-indole-1-carboxamide

2 mL of acetone was added to 30.1 mg of 5-({2-[({4-[1 -(2-hydroxyethyl)piperidin-4-yl]phenyl}carbonyl)amino]pyridin-4-yl}oxy)- 6-(2-methoxyethoxy)-N-methyl-1H-indole-1-carboxamide described in Production Example 1-15, then methanesulfonic acid (4.0 pl) was added and stirred at room temperature for 4 days. The solid was collected by filtration to obtain the title compound (20.4 mg). The powder X-ray diffraction angles of the obtained compound are shown below. (20 ± 0.2°): 11.7°, 13.7°, 15.2°, 16.9°, 18.0°, 18.7°, 19.9°, 21.1°, 22.0° and 24.1°.

[Reference Example 4]

Preparation of a crystal of 5-({2-[({4-[1-(2-hydroxyethyl)piperidin-4-yl]phenyl}carbonyl)amino]pyridin-4-yl}oxy)-6 tosylate salt -(2-methoxyethoxy)-N-methyl-1H-indole-1-carboxamide

2 mL of acetone was added to 30.7 mg of 5-({2-[({4-[1-(2-hydroxyethyl)piperidin-4-yl]phenyl}carbonyl)amino]pyridin-4-yl}oxy)- 6-(2-methoxyethoxy)-N-methyl-1H-indole-1-carboxamide described in Production Example 1-15, then p-toluenesulfonic acid monohydrate (12.3 mg) was added and stirred at room temperature for 4 days. The solid was collected by filtration to obtain the title compound (15.1 mg). The powder X-ray diffraction angles of the obtained compound are shown below. (20 ± 0.2°): 11.9°, 12.6°, 13.5°, 13.8°, 17.6°, 18.0°, 18.6°, 20.4°, 21.4° and 23.3°.

[Reference Example 5]

Preparation of a crystal of 5-({2-[({4-[1-(2-hydroxyethyl)piperidin-4-yl]phenyl}carbonyl)amino]pyridin-4-yl}oxy)-6 benzoate salt -(2-methoxyethoxy)-N-methyl-1H-indole-1-carboxamide

0.2 ml of ethyl acetate was added to a mixture of 20.3 mg of 5-({2-[({4-[1-(2-hydroxyethyl)piperidin-4-yl]phenyl}carbonyl)amino]pyridin-4- yl}oxy)-6-(2-methoxyethoxy)-N-methyl-1H-indole-1-carboxamide described in Production Example 1-15 and benzoic acid (8.91 mg) and stirred at room temperature. After 2 hours, 0.1 ml of ethyl acetate was added, and the reaction mixture was further stirred overnight. The solid was collected by filtration to obtain the title compound. The powder X-ray diffraction angles of the obtained compound are shown below. (20 ± 0.2°): 9.3°, 13.9°, 14.5°, 15.8°, 18.1°, 19.4°, 20.5°, 21.3°, 22.6° and 26.2°.

[Reference Example 6]

Preparation of a crystal of 5-({2-[({4-[1-(2-hydroxyethyl)piperidin-4-yl]phenyl}carbonyl)amino]pyridin-4-yl}oxy)-6 fumarate salt -(2-methoxyethoxy)-N-methyl-1H-indole-1-carboxamide

2 mL of acetone was added to a mixture of 30.6 mg of 5-({2-[({4-[1-(2-hydroxyethyl)piperidin-4-yl]phenyl}carbonyl)amino]pyridin-4-yl} oxy)-6-(2-methoxyethoxy)-N-methyl-1H-indole-1-carboxamide described in Production Example 1-15 and fumaric acid (7.24 mg), and stirred at room temperature overnight. The solid was collected by filtration to obtain the title compound. The powder X-ray diffraction angles of the obtained compound are shown below. (20 ± 0.2°): 9.6°, 13.8°, 15.7°, 16.7°, 19.8°, 21.0°, 22.0°, 22.4°, 24.7° and 25.7°.

[Reference Example 7]

Preparation of a crystal of 5-({2-[({4-[1(2-hydroxyethyl)piperidin-4-l]phenyl}carbonyl)amino]pyridin-4-yl}oxy)-6- (2-methoxyethoxy)-N-methyl-1H-indole-1-carboxamide

2 ml of acetone and 6N hydrochloric acid (10.0 pl) were added to 29.5 ml of 5-({2-[({4-[1-(2-hydroxyethyl)piperidin-4-yl]phenyl}carbonyl)amino] pyridin-4-yl}oxy)-6-(2-methoxyethoxy)-N-methyl-1H-indole-1-carboxamide described in Production Example 1-15, and the reaction mixture was stirred at room temperature. The title compound was isolated from the solvent as an oil.

[Reference Example 8]

Preparation of a crystal of 5-({2-[({4-[1-(2-hydroxyethyl)piperidin-4-yl]phenyl}carbonyl)amino]pyridin-4-yl}oxy)-6 hydrobromide salt -(2-methoxyethoxy)-N-methyl-1H-indole-1-carboxamide

2 ml of acetone and hydrobromic acid (7.8 pl) were added to 32.7 mg of 5-({2-[({4-[1-(2-hydroxyethyl)piperidin-4-yl]phenyl}carbonyl)amino]pyridin- 4-yl}oxy)-6-(2-methoxyethoxy)-N-methyl-1H-indole-1-carboxamide described in Production Example 1-15, and the reaction mixture was stirred at room temperature. The title compound was isolated from the solvent as an oil.

[Essay example]

The following Test Examples were carried out, and the physical properties or pharmacological effects of the compound (I) described in Production Example 1-15 or the salts of the compound (I) and their crystals were evaluated.

[Test Example 1] Hygroscopic nature

A dynamic vapor sorption apparatus was used to evaluate the hygroscopic nature of the 1.5 succinate salt of compound (I) of Example 1. The temperature of the sample forming part of the apparatus was maintained at 25°C and the relative humidity was adjusted (HR) step by step within a range of 5% to 95%. Relative humidity was regulated by adjusting the relative flow rates of dry nitrogen at 0% RH and 100% humidity. The weight of the sample was measured every 2 minutes with the microbalance. Moisture was successively changed when the magnitude of weight change over 5 minutes was less than 0.01%. The result is shown in Figure 5.

[Test Example 2] Cell-free kinase inhibitory activity

To a white flat bottom 96-well plate (Sumitomo Bakelite Co., Ltd., MS-8496W), 10 gl of FGFR1 protein solution (Carna Biosciences, Inc., 08-133) diluted to 1 gg/ml was added. with assay buffer (20 mM HEPES-NaOH, 0.01% Triton X-100, 2 mM DTT and 5 mM MgCl ² ), 10 gl of an assay buffer containing CSK-tide substrate (Ana Spec Inc., 63843 ) in a final concentration of 1000 nM and ATP (Promega Corporation, V9102) in a final concentration of 58.3 gM, and 5 gl of a test substance diluted with assay buffer, and the reaction was carried out at room temperature for 1 hour. To measure kinase activity, the ADP-Glo (TM) Kinase Assay (Promega Corporation, V9102) was used. After the reaction, 25 μl of ADP-Glo reagent was added to each well of the plate, and the reaction was carried out at room temperature for 40 minutes to stop the kinase reaction and to remove remaining ATP. Kinase detection reagent was additionally added, and the reaction was carried out at room temperature for 40 minutes, to cause the conversion of ADP to ATP, a luciferase/luciferin coupling reaction, and a light reaction by means of ATP. . To evaluate enzyme activity, the amount of luminescence in each well was measured by Envision (TM) (PerkinElmer Co., Ltd.). Luminescence values of wells containing protein kinase without addition of test substance were defined as 100% and luminescence values of wells without addition of test substance and protein kinase were defined as 0. %. Then, the ratio of luminescence value in the presence of the test substance was calculated. On the basis of this luminescence value ratio, the concentration of the test substance required to inhibit the kinase activity by 50% (ie, an IC ⁵⁰ value) was calculated.

FGFR2 cellular free kinase inhibitory activity, FGFR3 cellular free kinase inhibitory activity and FGFR4 cellular free kinase inhibitory activity were measured, respectively, using FGFR2 protein (Carna Biosciences, Inc., 08-134), FGFR3 protein (Carna Biosciences, Inc., 08-135) or FGFR4 protein (Carna Bioscience, Inc., 08-136) in the same manner as the case of the aforementioned FGFR1 cellular free kinase inhibitory activity. However, with respect to ATP concentration, the inhibitory activity of cellular free kinase was evaluated at a final concentration of 35 gM for FGFR2, at a final concentration of 16.7 gM for FGFR3, and at a final concentration of 75 gM for FGFR4. For FGFR3 and FGFR4, the reaction with the test substance was carried out at room temperature for 2 hours. Their results are shown in Table 1.

<Cellular Free Kinase Inhibitory Activity Data>

[Test Example 3] SNU-16 growth inhibition assay

The human stomach cancer cell line SNU-16 (ATCC Number CRL-5974) has been reported to harbor amplification of the FGFR2 gene (Cancer Res. 2008. 68: 2340-2348). SNU-16 cells were maintained in RPMI-1640 medium (Wako Pure Chemical Industries, Ltd., 187-02021) containing 10% FBS, and penicillin/streptomycin (Wako Pure Chemical Industries, Ltd., 168-23191). in a 5% CO ² (37°C) incubator. To each well of a 96-well plate (Becton, Dickinson and Company, 35-3075), 150 gl of SNU-16 cell suspension adjusted to a concentration of 1 x 104 cells/ml with RPMI-1640 medium containing 10% FBS, and the cell was incubated overnight in a 5% CO 2 incubator (37°C). On the next day, 50 gl of a test substance with RPMI-1640 medium containing 10% FBS was added, and the resulting part was incubated for 3 days in a 5% CO 2 incubator (37°C). Subsequently, 10 gl of Cell Counting Kit-8 (Dojindo Laboratories, CK04) was added to each well, and the resulting portion was incubated for 1 hour to 2 hours in a 5% CO ² incubator (37°C) to cause a color reaction. Absorbance value was measured with ENVISION (PerkinElmer Co., Ltd.) at 450 nm. The absorbance value of the wells without adding the test substance was defined as 100% and the absorbance value of the wells containing no cells was defined as 0%. Then, the absorbance ratio in the presence of the test substance was calculated. The concentration of the test substance required to inhibit cell growth by 50% (ie, an IC ⁵⁰ value) was calculated, and is shown in Table 2.

<SNU-16 growth inhibitory activity evaluation data>

[Test Example 4] Antitumor effect in SNU-16 subcutaneous xenograft model in mice

A human stomach cancer cell line SNU-16, which had been cultured in RPMI-1640 medium containing 10% FBS, and penicillin/streptomycin at a concentration of 1 x 108 cells/mL, was set. with Hank's Balanced Salt Solution (GIBCO #24020) to prepare a cell suspension, and the suspension was mixed with MATRlGEL (BD Biosciences, Cat# 354234) in a ratio of 1:1 to prepare a cell suspension at a concentration of 5 x 107 cells/mL. The cell suspension in a volume of 100 µl was inoculated into a subcutaneous part of the right flank of a nude mouse, 6 to 7 weeks old (BALB/cAJcl-nu/nu, female, Clea Japan Inc.). Seven days after the cell inoculum, the shortest and longest diameter of the tumor in each mouse were measured using an electronic digital caliper (Digimatic TM caliper, Mitutoyo Corporation) to calculate the tumor volume according to the following calculation formula:

Tumor volume (mm3) = Longest diameter (mm)

x Shortest diameter (mm) x Shortest diameter (mm) / 2

Based on tumor volumes obtained on the first day of administration, nude mice were grouped such that mean tumor volumes were substantially equal between groups. Each test substance was dissolved in DMSO, Tween 80 was added thereto to prepare a solution of 10-fold concentration, and the thus prepared solution was stored in a freezer before use. Immediately prior to administration, the stock solution was diluted with 5% glucose solution to obtain a final administration solution (in which the % ratio of DMSO, Tween 80, and 5% glucose solution was 3.5). :6.5:90). Each evaluation sample was orally administered to the test substance administration group at a volume of 20 ml/kg once a day continuously for 11 days, and in a control group, an administration solvent was administered. orally under the same conditions. Incidentally, the experiment was carried out on groups consisting of 5 mice.

With respect to each of the control groups and the test substance administration group, the ratio of the weight measured on the final day to the weight measured on the first day (relative body weight: RBW) was calculated. If the ratio of RBW of test substance administration group/RBW of control group is 0.9 or more, the corresponding test substance administration group was defined as well tolerated. In the test substance administration group thus defined as well tolerated, a tumor volume ratio of the test substance dosage group to the tumor volume of the control group obtained on the last day (T/ C) (%), and is shown in Table 3.

<Evaluation data of antitumor effect in SNU-16 subcutaneous xenograft model in mice>

Tl 1

Claims (12)

1. A salt consisting of 5-({2-[({4-[1-(2-hydroxyethyl)piperidin-4-yl]phenyl}carbonyl)amino]pyridin-4-yl}oxy)-6-( 2-methoxyethoxy)-N-methyl-1H-indole-1-carboxamide represented by formula (I):

and succinic acid or maleic acid. 2. The salt according to claim 1, which is a succinate salt. 3. The salt according to claim 2, which is 1.5 5-({2-[({4-[1-(2-hydroxyethyl)piperidin-4-yl]phenyl}carbonyl)amino]pyridin-4- succinate salt yl}oxy)-6-(2-methoxyethoxy)-N-methyl-1 H -indole-1-carboxamide. 4. The salt according to claim 2, which is 0.5 5-({2-[({4-[1-(2-hydroxyethyl)piperidin-4-yl]phenyl}carbonyl)amino]pyridin-4- succinate salt yl}oxy)-6-(2-methoxyethoxy)-N-methyl-1 H -indole-1-carboxamide. 5. The salt according to claim 1, which is a 5-({2-[({4-[1-(2-hydroxyethyl)piperidin-4-yl]phenyl}carbonyl)amino]pyridine-4 maleate salt -yl}oxy)-6-(2-methoxyethoxy)-N-methyl-1 H -indole-1-carboxamide. 6. A crystal of 5-({2-[({4-[1-(2-hydroxyethyl)piperidin-4-yl]phenyl}carbonyl)amino]pyridin-4-yl}oxy)- succinate salt 1.5 6-(2-methoxyethoxy)-N-methyl-1H-indole-1-carboxamide, which has diffraction peaks at (20 ± 0.2°) diffraction angles of 22.4°, 25.3° and 23.3° in X-ray diffraction X in powder form measured with copper irradiation; detector: scintillation counter; tube voltage: 50kV; tube current: 300mA; Slit: 0.5mm divergence slit (2mm height limitation slit), open spreading slit, open receiving slit; sweep speed: 10°/min; sampling interval: 0.02°; sweep range: 5° to 35°; sample holder made of aluminium. 7. A crystal (a) of 0.5 5-({2-[({4-[1-(2-hydroxyethyl)piperidin-4-yl]phenyl}carbonyl)amino]pyridin-4-yl} succinate salt oxy)-6-(2-methoxyethoxy)-N-methyl-1 H -indole-1 -carboxamide, having diffraction peaks at diffraction angles (20 ± 0.2°) of 19.8°, 15.7° and 13.9° in diffraction X-rays in powder form measured with copper irradiation; detector: scintillation counter; tube voltage: 50kV; tube current: 300mA; Slit: 0.5mm divergence slit (2mm height limitation slit), open spreading slit, open receiving slit; sweep speed: 10°/min; sampling interval: 0.02°; sweep range: 5° to 35°; sample holder made of aluminium. 8. A crystal of 5-({2-[({4-[1-(2-hydroxyethyl)piperidin-4-yl]phenyl}carbonyl)amino]pyridin-4-yl}oxy)-6 maleate salt -(2-methoxyethoxy)-N-methyl-1H-indole-1-carboxamide, having diffraction peaks at diffraction angles (20 ± 0.2°) of 20.1°, 17.0° and 16.2° in X-ray diffraction in powder form measured with copper irradiation; detector: scintillation counter; tube voltage: 50kV; tube current: 300mA; Slit: 0.5mm divergence slit (2mm height limitation slit), open spreading slit, open receiving slit; sweep speed: 10°/min; sampling interval: 0.02°; sweep range: 5° to 35°; sample holder made of aluminium. 9. A crystal of 5-({2-[({4-[1-(2-hydroxyethyl)piperidin-4-yl]phenyl}carbonyl)amino]pyridin-4-yl}oxy)- succinate salt 1.5 6-(2-methoxyethoxy)-N-methyl-1H-indole-1-carboxamide, which has peaks at chemical shifts (±0.5 ppm) of 108.5 ppm, 155.1 ppm, and 179.9 ppm in a solid-state NMR spectrum of 1 *3C. 10. The crystal of claim 9, having peaks at chemical shifts (±0.5 ppm) of 27.1 ppm, 34.8 ppm, 108.5 ppm, 155.1 ppm and 179.9 ppm in a 13C solid state NMR spectrum. 11. A crystal of 5-({2-[({4-[1-(2-hydroxyethyl)piperidin-4-yl]phenyl}carbonyl)amino]pyridin-4-yl}oxy)- succinate salt 6-(2-methoxyethoxy)-N-methyl-1H-indole-1-carboxamide, measured with copper irradiation; detector: scintillation counter; tube voltage: 50kV; tube current: 300mA; slit: 0.5mm divergence slit (2mm height limitation slit), open spreading slit, open receiving slit; sweep speed: 10°/min; sampling interval: 0.02°; sweep range: 5° to 35°; sample holder made of aluminum and having an X-ray powder diffraction pattern shown in Figure 1. 12. A pharmaceutical composition comprising the salt or crystal according to any one of claims 1 to 11 as active ingredient.